The overall objective of this proposal is the elucidation of mechanisms involved in regulation of the transcription of specific genes in viral oncogenesis. Studies will focus on transciptional events during the productive infection of human KB cells by human adenovirus 2 with the following aims: (1) to identify the complement of DNA-dependent RNA polymerases involved (and to what extent) in the transcription of specific viral RNAs (including the 5.5S species) during infection by analysis of endogenous enzyme-template systems (isolated nuclei) in conjunction with specific RNA polymerase inhibitors; (2) to purify and compare the structural and functional properties of the relevant DNA-dependent RNA polymerases in control and infected cells; (3) to use defined in vitro systems for analysis of the intrinsic specificity of the RNA polymerases for recognizing specific initiation and termination sites on purified or partially-purified viral templates and for identification of other factors (either soluble or associated with the template or enzyme) which may be required for specificity. Initial studies will focus on transcription of the gene for the 5.5S viral-associated RNA (presumably via a distinct RNA polymerase) while later studies will emphasize transcription from other viral sequences (via host RNA polymerase II, or a modified form thereof). Specific analytical methods will include nucleic acid hydridization and partial nucleic acid sequence techniques. The ultimate goal is to extend these studies to analysis of transcriptional controls in the establishment and/or maintenance of the transformed state. BIBLIOGRAPHIC REFERENCES: R. Weinmann, T. Brendler, H.J. Raskas, and R.G. Roeder, Identification of Small Viral-Coded RNA's Transcribed by RNA Polymerase III in Adenovirus 2 Infected Cells, Cell, 7, 557-566 (1976). R. Weinmann, J.A. Jaehning, H.J. Raskas, and R.G. Roeder, Viral RNA Synthesis and Levels of DNA-dependent RNA Polymerases During Replication of Adenovirus 2, J. Virology, 17, 114-126 (1976).